NEED HELP with non-denaturing IEF

Aye M.Tin muamt2 at uxa.ecn.bgu.edu
Mon Sep 25 14:37:00 EST 1995


Hi,
I am a graduate student trying to run IEF gel under non-denaturing 
condition to visualize peroxidase isozyme bands from barley leaves. I've 
been using 5% acrylamide, 10% glycerol, 3% ampholytes for my gel. My 
samples are extracts of homogenized barley leaves ( I homogenized barley 
leaves in 0.01M sodium phosphate buffer, pH 6, centrifuged down and took 
supernatent). So far I only have two successful results that show well 
focused bands. After that, I have been getting a smear of stain (for 
enzyme activity) along the lane for my samples but protein standards 
produced bands.  I am at a loss now as to what to do next. If any one 
has any suggestion, please let me know. I would really appreciate it.
Sincerely,
Aye M. Tin





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